菊花绿变病植原体在中国的发生及分子鉴定

对内蒙古农业大学校园内表现花器绿变症状的菊花样品进行采集和DNA提取,应用植原体16S rRNA基因和rp基因的引物进行巢式PCR扩增,从感病样品中分别扩增得到了长度均约为1.2 kb的片段。序列一致性分析表明,菊花绿变植原体16S rRNA基因与翠菊黄化植原体匈牙利风信子株系(GenBank登录号MN080271)、印度玉米株系(KY565571)、印度繁缕株系(KC623537)和印度马铃薯株系(KC312703)的核酸一致性最高,为99.9%,rp基因序列与翠菊黄化植原体立陶宛洋葱株系(GU228514)的核酸一致性最高,为99.8%。基于16S rRNA基因和rp基因构建系统进化树时发现,菊花绿变植原体均与16SrI-B亚组成员聚为一起。16S rRNA基因相似性系数分析表明,菊花绿变植原体与洋葱黄化植原体(AP006628)的相似性系数最高为1.00,洋葱黄化植原体(AP006628)在分类上属于16SrI-B亚组。因此,我们可以确定该菊花绿变植原体属于16SrI-B亚组。这是我国首次报道菊花绿变病的发生。     

Mycobacteriosis caused by Mycobacterium marinum in reared mullets: first evidence from Sardinia (Italy)

Mycobacterium marinum is a slow‐growing non‐tuberculous mycobacterium, and it is considered the most common aetiologic agent of mycobacteriosis in wild and cultured fish. The diagnosis is principally made by histology when positive Ziehl–Neelsen stain granulomas are detected. The aim of this study was to investigate the occurrence of mycobacteriosis in extensively cultured Mugilidae of two lagoons (Cabras and San Teodoro) from Sardinia by the use of histology, microbiology, PCR and DNA sequencing. Nine of 106 mullets examined were affected by mycobacteriosis, and the spleen was the most affected organ. The histology detected higher rate (100%) of infection in spleen than the culture and PCR (75% and 62.5%, respectively). The sequencing of hsp65 gene identified M. marinum as the primary cause of mycobacteriosis in the mullets examined. Mullets affected by mycobacteriosis were mainly fished in the San Teodoro lagoon characterized by critical environmental conditions. Histology remains the most common method in detecting fish affected by mycobacteriosis, and PCR‐based methods are essential for species identification. Our finding are worthy of attention because mycobacteriosis caused by M. marinum in reared mullets was evidenced for the first time in Sardinia, suggesting that this disease may be underestimated also in other cultured fish species.     

二倍体和四倍体泥鳅线粒体ND-5/6基因多态性的PCR-RFLP分析

采用PCR-RFLP分析方法,对洞庭湖、武汉两地的二倍体和四倍体泥鳅线粒体DNAND-5/6基因多态性及4个群体遗传变异和遗传关系进行了研究。结果表明,120尾泥鳅mtDNAND-5/6扩增片段长度均为2.2kb;从13种限制性内切核酸酶中筛选出6种多态性内切酶(HaeⅢ、DraⅠ、RsaⅠ、TaqⅠ、HinfⅠ、MspⅠ),对扩增产物进行酶切,共检测到66种单倍型。泥鳅群体内单倍型多样性和核苷酸多样性分别为0.7679～0.9385和0.01041～0.03212;其中,洞庭湖二倍体核苷酸多样性最高(0.03212),武汉二倍体其次(0.02452),二倍体群体内核苷酸多样性高于四倍体。群体间的核苷酸多样性(π)大小为0.02588～0.04144,平均值为(0.031737±0.000005)。群体间的核苷酸歧化距离(δ)大小为0.00462～0.01617,平均值为(0.010659±0.000003);其中,武汉二倍体和四倍体之间的核苷酸歧化距离最大(0.01617),武汉四倍体和洞庭湖二倍体之间的核苷酸歧化距离最小(0.00462)。MonteCarlo模拟x2检验表明,4个群体间的单倍型频率存在极...     

Cryptic improvement for fertility by continuous selfing of diploid potatoes using Sli gene

While a diploid potato was continuously selfed by a function of Sli gene, the level of heterozygosity reduced, and the fertility also reduced (inbreeding depression), which might be improved because continuous selfing could eliminate undesirable recessive alleles. To explore what was occurring as advancing self-generations in terms of fertility, 51 plants of an S₃ diploid potato family were evaluated for fertility-related traits and analyzed by RFLP markers. The level of heterozygosity was positively correlated with any of fertility-related traits (r = 0.108−0.333). At least six loci on six chromosome sections were associated with fertility-related quantitative trait loci (QTLs), of which three performed better in the heterozygotes and three in the homozygotes. By further continued selfing, QTLs performing better in heterozygotes were likely fixed to homozygotes with secondly better performance, and those performing better in homozygotes were fixed to the best genotypes. Thus, the selfed progenies were cryptically being improved for fertility by genetic fixation to the best or better genotypes in fertility-related QTLs.     

Characterization of the A4 cytoplasmic male-sterile lines of sorghum using RFLP of mtDNA

Three sorghum cytoplasmic male sterile lines CSV4 A(V), CSV4 A(G₁) and CSV4 A(M), grouped as A₄, were compared with a milo (A₁) and two other non-milo (A₂ and A₃) cytoplasms for their RFLP patterns of mitochondrial DNA (mtDNA). A 9.7 kb clone from pearl millet mtDNA discriminated each of the three A₄ entries whereas other maize and pearl millet mtDNA clones used could not distinguish this group completely. The molecular differences within the A₄ cytoplasmic group offer some explanation for the inconsistency in the fertility restoration behaviour of these A₄ lines obtained with a definite set of testers in the field. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detection of linkage between RFLP markers and genes affecting anthocyanin pigmentation in maize (Zea mays L.)

Summary Linkages between molecular markers and genes involved in the expression of agronomical traits have already been described in all of the major crops. In most cases, the genetic model underlying the Quantitative Traits Loci (QTL) is discussed. Here, Restriction Fragment Length Polymorphisms (RFLPs) and Mapmaker-QTL have been used to pinpoint seven regions of the genome significantly correlated with four pigmentation qualitative traits of maize (Zea mays L.). Two of these, located on chromosomes 2 and 10, explain most of the variation of these traits. The R and B gene loci known to be involved in the regulation of the anthocyanin pathway map to the same regions and we suggest that these loci could be the candidate genes involved in the correlations detected with RFLPs. This type of result is in accordance with the hypothesis of the candidate gene which supposes that, if we have a very high density map of randomly-selected cDNA clones, it should theoretically be possible to associate a cloned genic sequence with a phenotypic trait where correlations are found.     

RFLP characterization of Indian Musa germplasm for clonal identification and classification

Summary Nineteen single-copy clones isolated from a PstI genomic library (cv. Maiden Plantain), and eight Vigna chloroplast DNA clones were used to probe total genomic DNA digests of 57 genotypes of Musa from India. The 19 genomic clones detected a total of 107 polymorphisms among the 57 genotypes. Principal coordinates and phenetic analyses of these data placed cultivars and species into distinct groups that were in general agreement with a previously published RAPD-based classification of these same plant materials. The 107 polymorphisms were sufficient to differentiate each clone from every other clone. Heterologous Vigna chloroplast DNA probes were used to characterize the cytoplasm of Musa cultivars and species. PCO analysis of these RFLPs were detected both within and between the generally recognized genome groups, indicating multiple hybridization pathways in the origin of hybrid clones. Data presented demonstrate that RFLPs are sufficiently abundant to classify Musa germplasm and that genetic relationships among Musa cultivars, based upon RFLP data, are in general agreement with relationships determined by analysis of morphology and RAPDs.     

Mitochondrial DNA Restriction Fragment Length Polymorphisms in Malus Species

Three apple (Malus×domestica) cultivars and 11 Malus accessions have been investigated by the probe hybridization method on their mitochondrial DNA (mtDNA). The gene probes used were: coxI, coxII, atpA, atp6, and atp9. Our results revealed enough variation to characterize ten mtDNA haplotypes among the Malus genotypes examined. The taxonomic and phylogenetic implications of mtDNA polymorphism are also discussed.     

Use of multiparental inbred populations to determine allelic relationships of molecular markers

The assessment of polymorphism exhibited by molecular markers is an arduous but essential task that facilitates the use of molecular tools by breeders and geneticists. For that purpose, the value of a wheat composite population was assessed for characterizing the diversity of restriction fragment length polymorphism (RFLP) markers developed by INRA-Génoblé. The polymorphism of 13 genomic probes was measured over a set of 80 inbred lines randomly extracted by single-seed descent from a composite-cross of 16 wheat lines. The 13 probéenzyme combinations revealed 27 loci with codominant polymorphism. As many bands were so far unmapped, the segregational analysis of the progenies appeared very suitable for complex patterns, both in determining allelic relationships and in revealing linkage between loci. Allelic diversity, band sizes and chromosomal location assessed from nullisomic-tetrasomic lines are given for the 27 loci.     

土壤微生物多样性的分子生态学研究方法

传统的平板培养法分离培养和鉴定土壤微生物只能反映极少数微生物的信息,种类只占土壤微生物种类总数的0.1%～1%,分子生态学方法应用于土壤微生物的鉴定显示出极大的优越性.着重阐述了土壤微生物多样性的研究内容、意义及目前的采用分子生态学的方法研究土壤微生物多样性,尤其以DGGE（denaturing gradient gel electrophoresis）分子生物学技术以及RAPD（Random amplified polymorphic DNA ）随机扩增的多态性分析方法更为精确和快速,为土壤微生物多样性研究提供了一个更加广阔的前景.